Calcium Signaling Protocols

ISBN-13: 9781493959280 / Angielski / Miękka / 2016 / 360 str.

David G. Lambert; Richard D. Rainbow

Opis

This book explores new protocols and features revised and improved methodologies, making it a valuable reference for all those contemplating a move into the study of intracellular Ca2+. It features lab-ready procedures and includes key implementation advice.

Spis treści

Part I: General

1. Fluorescent Measurement of [Ca²⁺]_c: Basic Practical Considerations

            Alec W.M. Simpson

2. Measurement of [Ca²⁺]_i in Whole Cell Suspensions Using Fura-2

            Anish Patel, Robert A. Hirst, Charlotte Harrison, Kazuyoshi Hirota, and David G. Lambert

Part II: Specialist Measurement Systems

3. Confocal Microscopy: Theory and Applications for Cellular Signaling

            Stephen C. Tovey, Paul J. Brighton, Edward T.W. Bampton, Yan Huang, and Gary B. Willars

4. Ratiometric Ca²⁺ Measurements Using the FlexStation^® Scanning Fluorometer

            Ian C.B. Marshall, Izzy Boyfield, and Shaun McNulty

5. Measuring Ca²⁺ Changes in Multiwell Format Using the Fluorometric Imaging Plate Reader (FLIPR^®)

            Ian C.B. Marshall, Davina E. Owen, and Shaun McNulty

6. Ratiometric [Ca²⁺]_i Measurements in Adherent Cell-lines Using the NOVOstar Microplate Reader

            Benjamin D. Hunt and David G. Lambert

Part III: Measurement of Ca²⁺ Channel Activity

7. Whole-Cell Patch Clamp Recording of Voltage-Sensitive Ca²⁺ Channel Currents in Single Cells: Heterologous Expression Systems and Neurones

            Jon Brown, Atticus H. Hainsworth, Alessandro Stefani, and Andrew D. Randall

8. Combined Calcium Fluorescence Recording with Ionic Currents in Contractile Cells

            Richard D. Rainbow

Part IV: Measurement of Ins(1,4,5)P₃ and Ca²⁺ Release Form Intracellular Stores

9. Measurement of Phospholipase C by Monitoring Inositol Phosphates Using [³H]Inositol Labeling Protocols in Permeabilized Cells

            Alison Skippen, Philip Swigart, and Shamshad Cockcroft

 

10. Single Cell Imaging Techniques for the Real-Time Detection of IP₃ in Live Cells

            Carl P. Nelson

11. Measurement of Inositol(1,4,5)Trisphosphate Using a Stereospecific Radioreceptor Mass Assay

            Darren Smart

Part V: Specialist Measurement Techniques

12. Measurement of [Ca²⁺]_i in Smooth Muscle Strips Using Front-Surface Fluorimetry

            Hideo Kanaide and Katsuya Hirano

13. Calcium Measurements from Whole Heart Using Rhod-2

            Bum-Rak Choi

14. Measurement of Changes in Endothelial and Smooth Muscle Ca²⁺ in Pressurized Arteries

            Kim A. Dora and Michael A. Hill

15. Single Cell and Subcellular Measurements of Intracellular Ca²⁺ Concentration

            John G. McCarron, Marnie L. Olson, Susan Chalmers, and John M. Girkin

16. Simultaneous Analysis of Intracellular pH and Ca²⁺ from Cell Populations

            Raul Martinez-Zaguilan, Linda S. Tompkins, Robert J. Gillies, and Ronald M. Lynch

17. Measurements of Ca²⁺ Concentration with Recombinant Targeted Luminescent Probes

            Denis Ottolini,Tito Calì, and Marisa Brini

18. Chimeric G-Proteins in Fluorimetric Calcium Assays: Experience with Opioid Receptors

            Valeria Camarda and Girolamo Calo

19. Compartmentalizing Genetically-Encoded Calcium Sensors

            David A. Williams, Mastura Monif, and Kate L. Richardson

20. Measurement of Cytosolic-Free Ca²⁺ in Plant Tissue

            Martin R. McAinsh and Carl K.-Y. Ng

21. Measurement of Ca²⁺-ATPase Activity (in PMCA and SERCA1)            Danuta Kosk-Kosicka

Blurb

The regulation of intracellular Ca²⁺ has continued to be a powerful area of study since the publication of the first and second editions of Calcium Signaling Protocols, and the developments in the field have also, naturally, continued.  With the third edition, expert contributors explore some of the exciting new molecular techniques that have both enabled new studies of intracellular Ca²⁺ regulation and provided much new information on processes.  Comprised of five main section, the book covers theoretical and very simple suspension-based fluorimetric assays, specialist measurement systems, measurement of channel activity, measurement of store release, as well as specialist measurement techniques which include targeted probes, using G-protein chimeras to force Ca²⁺ signalling for screening, and genetically encoded sensors.  Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

 

Updated and accessible, Calcium Signaling Protocols, Third Edition will be a valuable reference for all those contemplating a move into the study of intracellular Ca²⁺.